Here we plot the results of the metabolisation assay in liquid cultures of Microbacteria from the maize root bacteria (Thoenen et al. 2023) and the AtSphere collection (Bai et al. 2015). We tested MBOA and DIMBOA-Glc. Additionally we also report growth data of all Microbacteria in MBOA-containing TSB medium to test them for MBOA tolerance and in MBOA-containing minimal medium to test them for using MBOA as sole carbon source for growth.

MBOA metabolisation

Quantification & calculations: using a standard curve the samples are quantified

Explore raw data

Stacked Bargraph

Phylogenetic tree

DIMBOA-Glc metabolisation

Quantification & calculations using a standard curve the samples are quantified

join 11 stds and HMPAA

remove the compounds which are not reliable quantified

Dilutions: 150 + 350 = 500 50 + 700 = 750 500 / 150 = 3.33333 750 / 50 = 15

1/((500/150)*15) = 0.02

for some reason the MBOA concentrations are a bit too high (factor 1.273141 controlled with t0 MBOA sample) –> ev because of precipitaion issue?

Explore raw data

Phylogenetic tree

Metabolite categories for comparative genomics

MBOA

MBOA metabolisation
max_MBOA max_AMPO max_HMPAA
516.2433 302.6399 44.06674

DIMBOA-Glc metabolization

DIMBOA-Glc metabolisation
max_DIMBOA_Glc max_MBOA max_AMPO
575.2392 120.9501 7.355668

Groups

GROWTH MICROBACTERIA: TSB (Metabolite samples) & MM (Carbon source)

2nd run was left for 1 cycle longer, therefore the same amount of cycles was kept

Growth curves of all plates single treatments

Calculate AUC

To quantify the total bacterial growth over time, we calculate the total area under the curve. This is done with the function “auc()” For comparison between treatments, the total AUC (calculated using density increase) and AUC_raw (calculated using raw density) is normalized with the AUC of the strain grown in the control treatment (no chemicals added, just normal growth media with DMSO).

Plotting AUC

TSB and Minimal medium

Heatmap growth in minimal media

TSB: MBOA tolerance

Calculate growth reduction in MM MBOA compared to MM DMSO

Statistics MBOA tolerance in MM

Phylogenetic tree

COMBINED TREE

Thresholds for qualitative analysis - weak MBOA-degraders: >30% of MBOA degraded compared to the control) - strong MBOA-degraders: (>90% of MBOA degraded, Fig. 3a). strong AMPO-formers (100 - 10 %) weak AMPO-formers’, <10% of max. AMPO forming strain) (0.01 µM, limit of detection).

Tree with AMPO Phenotype on plates

Tree as above with MBOA metabolites

Tree as above with DIMBOA-Glc metabolites

## R version 4.3.1 (2023-06-16)
## Platform: x86_64-apple-darwin20 (64-bit)
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